Browse
Submit Data
Databases
API
Help

Dataset Information

321 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Time-dependent, Glucose-regulated Arabidopsis Regulator of G-Protein Signaling 1 Network

ABSTRACT: Plants lack 7-transmembrane, G-protein coupled receptors (GPCRs) because the G alpha subunit of the heterotrimeric G protein complex is “self-activating” - meaning that it spontaneously exchanges bound GDP for GTP without the need of a GPCR. In lieu of GPCRs, most plants have a seven transmembrane receptor-like Regulator of G Signaling (RGS) protein, a component of the complex that keeps G signaling in its non-activated state. The addition of glucose physically uncouples AtRGS1 from the complex through specific endocytosis leaving the activated G protein at the plasma membrane. Here, the complement of proteins in the AtRGS1/G-protein complex over time from glucose-induced endocytosis was profiled by immunoprecipitation coupled to mass spectrometry (IP-MS). A total of 119 proteins in the AtRGS1 complex were identified. Several known interactors of the complex were identified, thus validating the approach, but the vast majority were not known previously. AtRGS1 protein interactions were dynamically modulated by D-glucose. In response to D-glucose, a diverse range of proteins became maximally associated with AtRGS1, whereas endosomal trafficking proteins were more distinctively associated with AtRGS1 following D-glucose treatment. Functional analyses are consistent with core proteins operating in response to stimulus and other metabolic pathways and thus provide an insight into spatiotemporal and mechanistic aspects of AtRGS1 regulation.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Root, Shoot

SUBMITTER: Evan McConnell

LAB HEAD: Leslie M. Hicks

PROVIDER: PXD003103 | Pride | 2016-06-15

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Similar Datasets

Spatial encoding of cyclic AMP signaling specificity by GPCR endocytosis

Project description:G protein-coupled receptors (GPCRs) are well known to signal via cyclic AMP (cAMP) production at the plasma membrane, but it is now clear that various GPCRs also signal after internalization. Apart from its temporal impact through prolonging the cellular response, we wondered if the endosome-initiated signal encodes any discrete spatial information. Using the beta2-adrenoceptor (β2-AR) as a model, we show that endocytosis is required for the full repertoire of downstream cAMP-dependent transcriptional control. Next, we describe an orthogonal optogenetic approach to definitively establish that the location of cAMP production is indeed the critical variable determining the transcriptional response. Finally, our results suggest that this spatial encoding scheme helps cells functionally discriminate chemically distinct β2-AR ligands according to differences in their ability to promote receptor endocytosis. These findings reveal a discrete principle for achieving cellular signalling specificity, based on endosome-mediated spatial encoding of intracellular second messenger production and 'location aware' downstream transcriptional control.

2014-11-01 | GSE57274 | GEO

Selection of the dimeric sweet taste receptor for surface expression and intersubunit interactions

Project description:The sweet taste receptor is a heterodimer of two class C G protein-coupled receptors (GPCRs), T1R2 and T1R3. While homologous structures of individual domains have been determined, the architecture of the full length T1R2-T1R3 dimer is unknown. Using random mutagenesis and cell sorting, we identify single point mutations and a C-terminal intracellular retention motif in human T1R2 that modulate surface expression and co-trafficking with T1R3 in a HEK293-derivative cell line. A comprehensive mutational scan of T1R2 based on surface expression of both subunits revealed conserved sites for T1R3 interactions, including surfaces on lobe 1 of the ligand binding domain, cysteine rich domain, and transmembrane helix 6. Using the mutational scan to guide modeling of the T1R2-T1R3 dimer predicts a twisted architecture that can explain how outwards motion of lobes 2 in the ligand-binding domains is translated into reorganization of transmembrane regions, and further predicts that extracellular loops 2 form continuous folded structures with the cysteine-rich domains near the dimer axis. These insights into the putative structural organization of the human sweet taste receptor have general implications for the mechanism of class C GPCRs.

2018-12-01 | GSE115751 | GEO

Human FZD6 phosphorylation LC-MS/MS

Project description:Frizzleds (FZD) function as receptors for secreted lipoglycoproteins of the Wingless/Int-1 (WNT) family, initiating an important signal transduction network in multicellular organisms. FZDs are classified as G protein-coupled receptors (GPCRs), which control a wide variety of physiological functions. GPCRs are well known to be regulated by phosphorylation, which can lead to specification of downstream signaling or receptor desensitization. The role and underlying mechanisms of phosphorylation of FZDs remains largely unexplored. Here, we investigated the phosphorylation of human FZD6. In order to assess FZD6 phosphorylation experimentally, we employed mass spectrometry. HEK293 cells expressing FZD6-GFP were treated with either control, WNT-3A or WNT-5A containing conditioned medium for 30 min and processed for LC-MS/MS analysis. Six phosphorylated serine residues were detected in the C-terminus of FZD6 − S592, S620, S629, S648, S653 and S656.

2018-10-18 | PXD011173 | Pride

Rod-derived Cone Viability Factor 2

Project description:Rod-derived Cone Viability Factor (RdCVF) is a truncated thioredoxin secreted by rod photoreceptors that protects cones. Because the secondary loss of cones in retinitis pigmentosa (RP) is the major visual handicap of this untreatable neurodegenerative disease, the administration of RdCVF is thought to be a promising therapy for RP. We show that RdCVF is acting by binding to Basigin-1 (BSG1) at the surface of cones. BSG1 is an alternative splice product of the BSG gene, a transmembrane protein with an extra immunoglobin domain expressed specifically in the retina. BSG1 binds to the glucose transporter GLUT1. RdCVF increases glucose entry into cones. We found that the increase in glucose is used by cones to induce cell survival by stimulating aerobic glycolysis. A disease associated missense mutation of RdCVF results in its inability to bind to BSG1, to stimulate glucose uptake and to prevent secondary cone death of a model of RP.

2016-06-15 | PXD001715 | Pride

Light effect via Opn3 on the activity of brown adipocytes

Project description:Opsin3 (Opn3) is a transmembrane heptahelical G-protein-coupled receptor (GPCR) with the potential to produce a non-visual photoreceptive effect. Interestingly, anatomical profiling of GPCRs reveals that Opn3 mRNA is highly expressed in adipose tissue. The photosensitive functions of Opn3 in mammals are poorly understood, and whether Opn3 has a function in fat is entirely unknown. In this study, we found that Opn3-knockout (Opn3-KO) mice were prone to diet-induced obesity and insulin-resistance. At the cellular level, Opn3-KO brown adipocytes cultured in darkness had decreased glucose uptake and lower nutrient-induced mitochondrial respiration than wild-type (WT) cells. Light exposure promoted mitochondrial activity and glucose uptake in WT adipocytes but not in Opn3-KO cells. Using RNA sequencing, we identified several novel light-sensitive and Opn3-dependent molecular signatures in brown adipocytes. Importantly, direct exposure of BAT to light in living mice significantly enhanced thermogenic capacity of BAT, and this effect was diminished in Opn3-KO animals. These results uncover a previously unrecognized cell-autonomous, light-sensing mechanism in brown adipocytes via Opn3-GPCR signaling that can regulate fuel metabolism and mitochondrial respiration in vitro and thermogenesis in vivo. Our work also provides a molecular basis for developing light-based treatments for obesity and its related metabolic disorders.

2020-02-26 | GSE140764 | GEO

Whey Protein-based Enteral Nutrition Support to Improve Protein Economy in Surgical Patients

Project description:A prospective, randomized and controlled study is proposed to establish whether an enteral nutrition support regimen based on pressurized whey protein and glucose improves the postoperative utilization of amino acid substrates compared to a drink based on glucose alone. The kinetics of protein metabolism (protein breakdown, protein synthesis and amino acid oxidation) will be investigated using stable isotope methodology before and after surgery in patients undergoing colon resection. Stable isotope infusions will be conducted one week before surgery and on the second postoperative day for two hours in the fasted state and for four hours while sipping the enteral nutrition support regimen. Patients will consume one of two enteral nutrition support regimens consisting of a drink containing either pressurized whey protein and glucose or glucose alone. It is hypothesized that an enteral nutrition support regimen based on pressurized whey protein and glucose promotes positive protein balance through increased protein synthesis or reduced protein breakdown compared to glucose alone.

| 2100821 | ecrin-mdr-crc

Mapping protein interactions of sodium channel NaV1.7 using epitope-tagged gene targeted mice

Project description:The voltage-gated sodium channel NaV1.7 plays a critical role in pain pathways. As well as action potential propagation, NaV1.7 regulates neurotransmitter release, integrates depolarizing inputs over long periods and regulates transcription. In order to better understand these functions, we generated an epitope-tagged NaV1.7 mouse that showed normal NaV1.7 channel activity and normal pain behavior. Analysis of NaV1.7 complexes affinity-purified under native conditions by mass spectrometry revealed 267 NaV1.7 associated proteins including known and novel interactors such as sodium channel β3 subunit (Scn3b) and collapsin response mediator protein (Crmp2). Selected NaV1.7 protein interactors, such as Crmp2, membrane-trafficking protein synapototagmin-2 (Syt2), G protein-regulated inducer of neurite outgrowth 1 (Gprin1), L-type amino acid transporter 1 (Lat1) and transmembrane P24 trafficking protein 10 (Tmed10) were validated using co-immunoprecipitation and functional assays. The information provided with this physiologically normal epitope-tagged mouse should provide useful insights into the downstream mechanisms associated with NaV1.7 channel function.

2018-01-15 | PXD005155 | Pride

Proteomic view of the Plasmodium falciparum life cycle

Project description:The completion of the Plasmodium falciparum clone 3D7 genome provides a basis on which to conduct comparative proteomics studies of this human pathogen. Here, we applied a high-throughput proteomics approach to identify new potential drug and vaccine targets and to better understand the biology of this complex protozoan parasite. We characterized four stages of the parasite life cycle (sporozoites, merozoites, trophozoites and gametocytes) by multidimensional protein identification technology. Functional profiling of over 2,400 proteins agreed with the physiology of each stage. Unexpectedly, the antigenically variant proteins of var and rif genes, defined as molecules on the surface of infected erythrocytes, were also largely expressed in sporozoites. The detection of chromosomal clusters encoding co-expressed proteins suggested a potential mechanism for controlling gene expression. Keywords: ordered

2002-12-22 | E-GEOD-92 | biostudies-arrayexpress

Proximity labeling of host factor ANXA3 in HCV infection reveals a novel LARP1 function in viral entry

Project description:Hepatitis C virus (HCV) infection is tightly connected to the lipid metabolism with lipid droplets (LDs) serving as assembly sites for progeny virions. A previous LD proteome analysis identified annexin A3 (ANXA3) as an important HCV host factor that is enriched at LDs in infected cells and required for HCV morphogenesis. To further characterize ANXA3 function in HCV, we performed proximity labeling using ANXA3-BioID2 as bait in HCV-infected cells. Two of the top proteins identified proximal to ANXA3 during HCV infection were the La-related protein 1 (LARP1) and the ADP ribosylation factor-like protein 8B (ARL8B), both of which have been previously described to act in HCV particle production. In follow-up experiments ARL8B functioned as a pro-viral HCV host factor without localizing to LDs und thus likely independent of ANXA3. In contrast, LARP1 interacts with HCV core protein in an RNA-dependent manner and is translocated to LDs by core. Knockdown of LARP1 decreased HCV spreading without altering HCV RNA replication or viral titers. Unexpectedly, entry of HCV particles and E1/E2-pseudotyped lentiviral particles was reduced by LARP1 depletion, whereas particle production was not altered. Using a recombinant vesicular stomatitis virus (VSV)ΔG entry assay, we showed that LARP1 depletion also decreased entry of VSV with VSV, MERS, and CHIKV glycoproteins. Therefore, our data expand the role of LARP1 as an HCV host factor that is most prominently involved in the early steps of infection, likely contributing to endocytosis of viral particles through the pleiotropic effect LARP1 has on the cellular translatome.

2024-04-17 | PXD049182 | Pride

Glucose dissociates DDX21 dimers to regulate mRNA splicing and promote epidermal differentiation

Project description:We discovered that glucose directly bind to DDX21, induced the conformation change of the proteins and inhibit its helicase activity. Glucose also inhibits the dimerization of DDX21, re-localize DDX21 from nucleolus to nucleoplasm and reassemble DDX21 to RNA splicing complex. DDX21 is essential for epidermal differentiation while with normal glucose condition, DDX21 was re-localized from nucleolus to nucleoplasm, bind to splicing factors and mRNA introns more to regulate splicing of key differentiation factors and promote epidermal differentiation.

2023-01-05 | PXD028561 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data